Ovarian Structures Research Articles

6615 Clinical features and genetic etiology of primary ovarian insufficiency in 46,XX children

Abstract Disclosure: F. Wang: None. X. Yin: None. X. Lan: None. P. Li: None. Purpose: The clinical features of primary ovarian insufficiency in 46,XX children were analyzed, the pathogenic genes of POI were screened, and the genetic etiology of primary ovarian insufficiency in children 46,XX was clarified. Method: The clinical data of 46,XX POI patients were collected, including uterine and ovarian ultrasound, serum follicle-stimulating hormone (FSH), estrogen, anti-mullerian hormone (AMH), chromosomal karyotype analysis, adrenal and thyroid function, and immune function. Whole exome sequencing (WES) and CNV were used to screen POI candidate genes. Novel mutations such as MCM8 gene were used to verify cell function. HE staining, immunohistochemistry, cell fluorescence, and Tunnel method were used to observe the changes in ovarian tissue structure. Construction overexpression plasmids of Mutant MCM8 gene in Hela cells. Result: 1. Clinical features of 46,XX POI: The age of onset of the children was 11.9∼16.1 years old. In addition to POI, the clinical phenotype showed height growth retardation. Among them, 4 cases were short stature, 3 cases were growth hormone deficiency, and 5 cases were backward in bone age. All of them showed elevated FSH (>25 IU/L), and 7 cases had low serum AMH (<0.5 ng/ml). 2.The Screening of POI candidate genes and pathogenicity analysis: 5 of the 10 children carried the mutations of the pathogenic genes with POI, that is, 2 cases (sisters of the same family) carried two novel heterozygous mutations of MCM8 gene, one infant carried two novel heterozygous mutations of CYP19A1 gene, and one case with delayed puberty carried one homozygous mutation of ZSW1M7 gene. 3.The MCM8 mutant were functionally validated: The sisters carried two novel mutations, namely p.C242R and p.S445*. The elder sister (the proband) showed POI, short stature and tumor susceptibility, while the younger sister showed a trend of gradual increase in FSH level and ovarian decline. The follicles of the proband disappeared and fibrous tissue proliferation, the MCM8 expression was lower, and the positive rate of cell apoptosis was higher than normal control. The downregulation of Bax and Capase3 expression in mutant MCM8 of Hela cell was delayed compared with wild type, indicating the persistence of apoptosis. Conclusion: 1. The clinical characteristics of 46,XX POI in children are ovarian hypoplasia, often combined with height growth retardation, biochemical manifestations of hypergonadotropic hypogonadism, decreased serum AMH, and karyotype 46,XX. 2.46,XX POI genetic etiology analysis found that five known POI-related pathogenic genes, namely CYP19A1, MCM8, and ZSW1M7, in which five novel mutations were identified.3.Mutant MCM8 promotes ovarian apoptosis by activating the Bcl-2/Bax/Caspase 3 pathway. Keywords: Primary ovarian insufficiency, short stature, apoptosis Presentation: 6/3/2024

Ovarian traits, spawning pattern and season, length at first maturity, and batch fecundity of Caragobius urolepis (Bleeker, 1852) caught from the Vietnamese Mekong Delta

Caragobius urolepis has a wide range of distribution, from brackish to freshwater water, ranging from India to the Philippines and Fiji, including the Vietnamese Mekong Delta (VMD), where it lives mainly in the coastal regions and plays a vital role in food supply. Our preliminary observation showed that its population tended to decline, but little is known about this species. This study, therefore, was conducted to provide data on the ovarian structure, spawning pattern and season, length at first maturity, and batch fecundity of this fish. The study of 306 specimens caught with trawl nets from April 2022 to March 2023 in two coastal areas of VMD—Dong Hai, Bac Lieu (DHBL), and Dam Doi, Ca Mau (DDCM)—showed that this goby spawned more than once because the oocytes in stage IV ovary showed different stages of development. It could release eggs monthly during a 12-month study, with a peak in July–September since stage IV ovaries were found monthly during the study period and the GSI exhibited a high value in July–September. The fish length at first maturity and batch fecundity varied with the site, as they were 18.8 cm and 3,757–9,187 (6,142 ± 707 SE eggs/female) in DDCM and 20.5 cm and 3,760–11,118 (5,634 ± 750 SE eggs/female) in DHBL. The findings provide valuable foundations for exploiting their resources and serve as a basis for studying artificial reproduction. It is suggested to avoid catching these fish species during the breeding season to ensure sustainable exploitation in the future.

Effect of ZnO NPs on Ovarian Histological Structure and Function in Adult Female Rats

تعد تقنية النانو واحدة من أهم التقنيات المستخدمة على نطاق واسع في العديد من المجالات من خلال تغيير خصائص المواد الفيزيائية والكيميائية لتكوين مواد جديدة تسمى الجسيمات النانوية لجعلها أكثر فعالية. تعتبرجسيمات أوكسيد الزنك النانوية هي واحدة من هذه الجزيئات التي تستخدم في تطبيقات متعددة ، خاصة في مستحضرات التجميل ومنتجات الوقاية من اشعة الشمس. ولهذا فإن استخدام هذه الجسيمات بكميات كبيرة يجعلها بتماس دائم مع الجسم والتي من الممكن أن تدخل الدورة الدموية بطرق مختلفة. كان الهدف من هذه الدراسة هو التحري عن تأثير جسيمات أوكسيد الزنك النانوي على التركيب النسجي والوظيفي للمبايض وكذلك تقييم مستوى هرمونات الغدد التناسلية (FSH و LH). ولاجل ذلك تم استخدام 54 من إناث الجرذان البالغة وقسمت بشكل عشوائي إلى ثلاث مجاميع رئيسية وفقًا الى فترات التجريع (1 ، 2 ، 4) أسابيع ، بعدها قسمت كل مجموعة رئيسية إلى ثلاث مجاميع فرعية ، كل مجموعة منها تتكون من 6 جرذان ، احدى هذه المجاميع تم استخدامها كمجموعة سيطرة وحقنت بـ 1 مل من الماء المقطر ، بينما المجموعتين الأخرى استخدمت كمجاميع معالجة وحقنت بـ 1 مل من محلول اوكسيد الزنك النانوي (ZnO NPs) بتركيزين مختلفين منخفض وعالي (50 و 200) مغم / كغم على التوالي ، بمعدل ثلاث جرعات في الأسبوع عن طريق الحقن داخل الصفاق. أظهرت النتائج تغيرات نسجية مختلفة في مقاطع أنسجة المبايض لدى الجرذان المجرعة بتراكيز وفترات زمنية مختلفة ، وكذلك أظهرت النتائج ايضا" انخفاض معنوي (p≤ 0.01) في مستويات هرمون الاستروجين والبروجسترون و LH و FSH ، لدى الحيوانات المعالجة بتراكيز وفترات زمنية مختلفة. وقد تم الأستنتاج بأن لجزيئات دقائق الزنك النانوية القدرة على إتلاف أنسجة المبيض ، وتعطيل وظائفه الخلوية مما أدى الى خلل في مستويات الهرمونات التناسلية (P و E2 و LH و FSH).

Microbiota profile in organs of the horseflies (Diptera: Tabanidae) in Northeastern China.

Tabanids, commonly known as horseflies and belonging to the family Tabanidae, are blood-feeding arthropods (BFA) found worldwide. They are known for their ability to mechanically and biologically transmit various animal pathogens. Tabanids are potential vectors for diseases such as Francisella tularensis, Anaplasma marginale, Theileria spp., and contributors to lumpy skin diseases. Despite their involvement in common BFA studies, tabanids have not been extensively explored in microbiome research. In this study, the microbiota structure and composition in various organs of four distinct genera of tabanids: Atylotus, Haematopota, Tabanus, and Hybomitra were examined. High-throughput sequencing of the bacterial 16S rRNA gene was performed to gain insights into the microbial communities associated with the different tabanid species. Result display that microbiota composition and diversity, including Firmicutes, Proteobacteria, and Bacteroidetes, varied significantly among the different organs, with the ovaries exhibiting significantly higher diversity. Apart from the Haematopota genus, Tenericutes were enriched in the midgut of other tabanid species, whereas the Malpighian tubules exhibited a higher abundance of Bacteroides. Notably, the ovarian microbiota structure was conserved among the four tabanid species, indicating its potential association with reproductive development. Evaluation of the potential pathogen risk revealed putative pathogens in over 100 genera associated with these tabanid commensal organisms. Twenty genera were annotated as zoonotic agents with a high abundance of Citrobacter and Brucella, highlighting the presence of this important group of zoonotic pathogens. Functional predictions of vector-microbiota interactions indicate that microbiota significantly affects vector biological traits and can influence pathogen transmission via direct interactions or by regulating host immunity and nutrition. For the first time, the distribution characteristics and functions of four genera of horsefly microbiota were analyzed, revealing the presence of multiple potential pathogenic microorganisms. These findings provide valuable insights for future research and the development of symbiotic-based strategies to control insect-borne diseases among tabanids.

Neuregulin 1 Signaling Attenuates Tumor Necrosis Factor α-Induced Female Rat Luteal Cell Death.

The corpus luteum (CL) is a transient ovarian endocrine structure that maintains pregnancy in primates during the first trimester and in rodents during the entire pregnancy by producing steroid hormone progesterone (P4). CL lifespan, growth, and differentiation are tightly regulated by survival and cell death signals through luteotrophic and luteolytic factors, including the epidermal growth factor (EGF)-like factor family. Neuregulin 1 (NRG1), a member of the EGF family, mediates its effect through ErbB2/3 receptors. However, the functional role of NRG1 in luteal cells (LCs) is unknown. Thus, this study investigated the role of NRG1 and its molecular mechanism of action in rat LC. Our experimental results suggest a strong positive correlation between steroidogenic acute regulatory protein (StAR) and NRG1 expression in mid-CL and serum P4 and estrogen (E2) production. In contrast, there was a decrease in StAR and NRG1 expression and P4 and E2 production with an increase in tumor necrosis factor α (TNFα) expression in regressing CL. Further in vitro studies in LCs showed that the knockdown of endogenous Nrg1 promoted the expression of proinflammatory and proapoptotic factors and decreased prosurvival factor expression. Subsequently, treatment with exogenous TNFα under these experimental conditions profoundly elevated proinflammatory and proapoptotic factors. Further analysis demonstrated that the phosphorylation status of ErbB2/3, PI3K, Ak strain transforming or protein kinase B (Akt), and ErK1/2 was significantly inhibited under these experimental conditions, whereas the treatment of TNFα further inhibited the phosphorylation of ErbB2/3, PI3K, Akt, and ErK1/2. Collectively, these studies provide new insights into the NRG1-mediated immunomodulatory and prosurvival role in LCs, which may maintain the function of CL.

Effect of repeated low doses of gonadotropin-releasing hormone on the secretion of luteinizing hormone and follicle-stimulating hormone, and ovarian function in dairy cows suffering from anovulation type I

The aim of the study was to evaluate the effect of repeated low doses of gonadotropin-releasing hormone (GnRH) agonist buserelin once a day for 5 days on follicle-stimulating hormone (FSH) and luteinizing hormone (LH) release, and ovarian function in dairy cows with anovulation type I. The study was conducted on 10 anovulatory Polish Holstein-Friesian cows. Cows in Group 1 (n = 5) received 4 µg of buserelin intramuscularly. once a day for 5 days. Control cows from Group 2 (n = 5) received saline. Concentrations of progesterone, FSH, and LH in the blood were analysed using radioimmunoassay (RIA). Ovarian structures were monitored weekly after the end of treatment by ultrasound for 4 weeks. Injections of buserelin increased FSH and LH concentrations. Release of FSH and LH was largely variable, but there was significant (P < 0.001) interaction between time and treatment during all treatment days. The number of follicles in the treatment group was significantly greater (P = 0.029) than in the control group on day 5, the diameter of follicles on day 12 was also larger (P < 0.01) in treated than in control cows. Ovulation occurred in 4 of the 5 treated cows, but only in 1 of the control cows in the 40 days after treatment. In conclusion, repeated low doses of GnRH analogue buserelin once a day for 5 days may increase the concentration of FSH and LH in dairy cows with anovulation type I.

Ruxolitinib Has a Protective Effect on Ovarian and Endometrial Tissues in Diabetic Rats via STAT3 Pathway

Background: Hyperglycemia is associated with ovarian dysfunction. Advanced glycation end products (AGE) may affect ovarian function by binding to particular AGE receptors (RAGE). Hematopoiesis and immunological conditioning are both controlled by the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. Many JAK-STAT signaling inhibitors, including ruxolitinib, have been approved to treat inflammatory disorders. We aimed to examine the potential protective effect of ruxolitinib, on ovarian dysfunction by comparing biochemical, pro-inflammatory, and histological abnormalities in a diabetic rat model. Methods: 24 female Wistar albino rats were included in the study. Diabetes was induced by streptozotocin (STZ) in 16 rats. Group 1: control (no diabetes mellitus, n = 8), Group 2 (diabetic = 8, 1 mL/kg/day saline, 4 weeks), and Group 3 (diabetic, n = 8, 2 mg/kg/day ruxolitinib, 4 weeks). The animals were euthanized, and bilateral hysterectomy and ovariectomy were performed for histopathological examination. The levels of signal transducer and activator of transcription 3 (STAT3) in tissue supernatants were measured. Results: Endometrial gland, ovarian stromal, and ovarian follicle degeneration scores were higher in group 2 compared with group 3 at p &lt; 0.001, whereas ovarian STAT3 level was significantly higher in group 2 compared with group 3 at p &lt; 0.001. Conclusions: Ruxolitinib can be a promising candidate for providing endometrial and ovarian structure continuity by JAK-STAT inhibition in diabetes.

Mechanism of Zuogui pill enhancing ovarian function and skin elastic repair in premature aging rats based on artificial intelligence medical image analysis.

AI medical image analysis shows potential applications in research on premature aging and skin. The purpose of this study was to explore the mechanism of the Zuogui pill based on artificial intelligence medical image analysis on ovarian function enhancement and skin elasticity repair in rats with premature aging. The premature aging rat model was established by using an experimental animal model. Then Zuogui pills were injected into the rats with premature aging, and the images were detected by an optical microscope. Then, through the analysis of artificial intelligence medical images, the image data is analyzed to evaluate the indicators of ovarian function. Through optical microscope image detection, we observed that the Zuogui pill played an active role in repairing ovarian tissue structure and increasing the number of follicles in mice, and Zuogui pill also significantly increased the level of progesterone in the blood of mice. Most of the ZGP-induced outcomes are significantly dose-dependent.

UNILATERAL OVARIAN TORSION IN FIVE GECKOS: CLINICAL AND ULTRASOUND FINDINGS.

Few cases of antemortem ovarian torsion and diagnosis have been described in reptiles. This case series reports clinical and ultrasound findings in five adult (aged 1-6 yr) female geckos (three leopard geckos [Eublepharis macularius], one crested gecko [Correlophus ciliatus], and one gargoyle gecko [Rhacodactylus auriculatus]) diagnosed with unilateral ovarian torsion between 2019 and 2023. All animals presented with acute weakness associated with coelomic distension, and one suffered from chronic diarrhea and cachexia. Coelomic ultrasound examination (12-MHz linear probe) revealed signs of bilateral follicular stasis and oophoritis in all cases (heterogenic follicles ≤1.3 cm diameter) associated with a large anechoic periovarian rim and a hyperechoic chord, consistent with twisted ovarian vessels. Blood supply to ovarian structures was not detected using a Doppler flow, and a unilateral ovarian torsion was diagnosed in all geckos. A bilateral ovariectomy was performed under general anesthesia in all five animals. The mass of the excised ovaries varied between 7 to 15 g (12.7-22.2% of body weight). One gecko died 1 d postsurgery; the four remaining animals were healthy 6 mon postsurgery. Gekkonids are unique among reptiles in that they undergo a monoautochronic ovulation (only one follicle is recruited by each ovary during each ovarian cycle); the presence of multiple vitellogenic follicles on each ovary facilitates the diagnosis of follicular stasis. This condition was present in all five geckos and was suspected to have led to ovarian torsion. This case series emphasizes the value of ultrasound examination for antemortem diagnosis of reproductive disorders in reptiles.

Bisphenol A Disrupts Ribosome Function during Ovarian Development of Mice.

This study examines the impact of Bisphenol A (BPA), a prevalent environmental estrogenic toxicant, on the ovarian development of mice. Mice were exposed to varying BPA doses from in utero to postnatal stages, up to weaning (day 21, PND 21) and puberty (day 45, PND 45). The BPA content in the serum of the offspring mice on PND 45 was higher than that of the mice sacrificed at PND 21. However, the ovary organ index of the mice of PND 21 was significantly increased, and the ovarian structure was damaged when exposed to BPA. In contrast, the mice with PND 45 did not show apparent ovarian lesions. On the other hand, granulosa cell apoptosis was detected in both PND 21 and PND 45 mice ovaries, and ERβ was increased under the influence of BPA. Transcriptomic analysis revealed BPA's significant impact on ribosomal gene expression, marked downregulation of Rpl21 and Rpsa, and upregulation of Rps2 in both age groups. These transcriptomic alterations were further corroborated by real-time PCR, highlighting a dose-dependent effect of BPA on Rps2. Our findings confirm BPA's detrimental effects on ovarian health, with more pronounced damage in younger mice, suggesting heightened vulnerability in this group. The study underscores ribosomes as critical targets in BPA-induced ovarian developmental disruptions.

HUMSCs restore ovarian function in POI mice by regulating GSK3β-mediated mitochondrial dynamic imbalances in theca cells

Premature ovarian insufficiency (POI), a major cause of female infertility, is defined as follicular atresia and a rapid loss of germ cells in women of reproductive age due to ovarian failure. Recently, findings from several studies have indicated that human umbilical cord mesenchymal stem cells (hUMSCs) can alleviate ovarian dysfunction resulting from POI. However, the mechanisms underlying this effect require further clarification. In this study, a mouse model of POI was established as achieved with an intraperitoneal injection of cyclophosphamide (CTX) into female C57BL/6J mice in vivo. These POI mice received a 1-week intervention of hUMACs. In addition, an in vitro POI model was also included. The cultured supernatants of hUMSCs and glycogen synthase kinase 3 beta (GSK3β) inhibitor (SB216763) were used to treat theca cells (TCs) exposed to CTX. Hematoxylin and Eosin (H&E) staining and Enzyme-linked immunosorbent assay (ELISA) were used to assess ovarian structure and morphology, as well as endocrine function in these POI mice. Based on results from the ELISA and JC-1 labeling, CTX exerted significant detrimental effects on testosterone levels and the mitochondrial membrane potential in TCs. Subsequently, Western Blot, Immunofluorescence staining (IF), and Quantitative real-time polymerase chain reaction (qRT-PCR) were used to evaluate various indicators of testosterone synthesis function and mitochondrial dynamics in ovaries and TCs of POI mice. In vivo, dysfunctions in ovarian structure and function in the POI mouse model were effectively restored following hUMSCs treatment, and abnormalities in hormone synthesis were significantly reduced. Furthermore, when the stem cell supernatants of hUMSCs were applied to TCs in vitro we found that GSK3β expression was reduced, the imbalance of mitochondrial dynamics was alleviated, and the ability of mitochondrial testosterone synthesis was increased. Taken together, our results indicate that hUMSCs treatment can restore the imbalance of mitochondrial dynamics and restart testosterone synthesis of TCs by suppressing GSK3β expression, ultimately alleviating POI damage.

The annual ovarian cycle in Northern pit vipers (Bothrops atrox) using serum steroids and ultrasound of the female reproductive tract

Knowledge on hormonal regulation of reproductive cycles in viperid snakes is still incipient, especially when it comes to females and tropical species. There is an urgent need to understand the reproduction of venomous snakes to improve assisted reproduction techniques and optimize the maintenance of these animals in captivity. With this in mind, we monitored Northern pit viper females year-round throughout different seasons via serum levels of progesterone (P4) and estradiol (E2) in conjunction with ultrasound examinations. Ovarian follicles were classified according to their size and stage of vitellogenesis in F-I and F-II (non-vitellogenic phase) or in F-III and F-IV (vitellogenic phase). During autumn and winter, five adult males were rotated among these females for reproductive pairing, which resulted in 17 copulations and 2 pregnancies in the first year and 12 copulations and 5 pregnancies in the second year. Then, we assessed changes in P4 and E2 levels according to seasons, predominant ovarian structures and the presence of embryos or eggs in the oviduct. Our findings showed high levels of E2 when a greater number of vitellogenic follicles were detected, indicating a possible influence of E2 on vitellogenesis and higher levels of P4 whenever eggs and embryos were visualized in the oviduct, implying its role in maintaining pregnancy. Descriptive analysis of the vipers’ ovarian cycles revealed a greater number of vitellogenic follicles during winter, probably as a result of increases in E2; whereas pregnancies occurred predominantly in spring, under the influence of P4. The use of ultrasound images, as a minimally invasive methodology, associated with serum steroid levels has proven to be an efficient approach in the reproductive monitoring of Northern pit vipers in vivo. In addition, these data suggest that female pit vipers under human care display a seasonal reproductive cycle, despite earlier studies involving captive males of the species indicating a lack of seasonality in sperm production and quality.

Bioengineered 3D ovarian model for long-term multiple development of preantral follicle: bridging the gap for poly(ε-caprolactone) (PCL)-based scaffold reproductive applications

BackgroundAssisted Reproductive Technologies (ARTs) have been validated in human and animal to solve reproductive problems such as infertility, aging, genetic selection/amplification and diseases. The persistent gap in ART biomedical applications lies in recapitulating the early stage of ovarian folliculogenesis, thus providing protocols to drive the large reserve of immature follicles towards the gonadotropin-dependent phase. Tissue engineering is becoming a concrete solution to potentially recapitulate ovarian structure, mostly relying on the use of autologous early follicles on natural or synthetic scaffolds. Based on these premises, the present study has been designed to validate the use of the ovarian bioinspired patterned electrospun fibrous scaffolds fabricated with poly(ε-caprolactone) (PCL) for multiple preantral (PA) follicle development.MethodsPA follicles isolated from lamb ovaries were cultured on PCL scaffold adopting a validated single-follicle protocol (Ctrl) or simulating a multiple-follicle condition by reproducing an artificial ovary engrafted with 5 or 10 PA (AO5PA and AO10PA). The incubations were protracted for 14 and 18 days before assessing scaffold-based microenvironment suitability to assist in vitro folliculogenesis (ivF) and oogenesis at morphological and functional level.ResultsThe ivF outcomes demonstrated that PCL-scaffolds generate an appropriate biomimetic ovarian microenvironment supporting the transition of multiple PA follicles towards early antral (EA) stage by supporting follicle growth and steroidogenic activation. PCL-multiple bioengineering ivF (AO10PA) performed in long term generated, in addition, the greatest percentage of highly specialized gametes by enhancing meiotic competence, large chromatin remodeling and parthenogenetic developmental competence.ConclusionsThe study showcased the proof of concept for a next-generation ART use of PCL-patterned scaffold aimed to generate transplantable artificial ovary engrafted with autologous early-stage follicles or to advance ivF technologies holding a 3D bioinspired matrix promoting a physiological long-term multiple PA follicle protocol.

Ultrasonographic threshold of ovarian structure in premenopausal women of different ethnicity

The polycystic ovarian morphology (PCOM) is a generally accepted ultrasound marker for ovulatory dysfunction, is one of the criteria for polycystic ovary syndrome (PCOS) and is established based on the assessment of ovarian volume (OV) and the follicle number per ovary (FNPO), taking into account the upper normal values determined in healthy premenopausal women. However, there is a necessity for regular revision of the PCOM characteristics depending on ethnic and age characteristics.The aim. To develop differentiated standards for assessing the ultrasonographic ovary structure in premenopausal women of various ethnicity.Materials and methods. From March 2016 to December 2019, a multicenter cross-sectional prospective study was conducted in Eastern Siberia (Irkutsk region) and in the neighboring Republic of Buryatia. The study included 1134 participants: 715 women of Caucasian origin, 312 Asian women, 107 women of mixed ethnic subpopulation.Results. It has been established that for Caucasians, it is advisable to diagnose PCOM when the ovarian volume is 9 cm3 and/or FNPO ≥ 12; for women of the Asian population – when the ovarian volume is 7 cm3 and/or FNPO ≥ 11; for women of mixed ethnicity – when the ovarian volume is 8 cm3 and/or FNPO ≥ 9. An important advantage of our study is that all participants were recruited from a non-selective multi-ethnic population of women with comparable socio-demographic characteristics living in the same geographical conditions.Conclusion. Differentiated approach for identifying the polycystic ovarian morphology in premenopausal women of different ethnic groups requires using ethnically differentiated normative readings

GC/MS Analysis and Compounds Isolation ofLycium shawii Petroleum Ether Seeds Extract for Regulating Nrf2/OH-1 Pathway, Oxidative Stress and Inflammation in Acrylamide- Induced Infertility in Female Rats.

Acrylamide is formed during heating of starchy foods at high temperature and induces reproductive toxicity. Our study is designed to evaluate the chemical constitution and anti-infertility effect of Lycium shawii seeds extract on female rats. Nutritional profile was estimated, and major active compounds were isolated and identified. Biological evaluation of Lycium shawii extract on female rats was performed and measured by prolactin, follicular stimulating hormone, luteinizing hormone, estradiol, progesterone, tumor necrosis factor-α, interleukin-6, heme oxygenase-1, nuclear respiratory factor-2, malondialdehyde, glutathione, DNA fragmentation and ovarian architecture parameters. Data revealed that presence of ɤ-tocopherol, vitamin C, magnesium and thirty-eight bioactive compounds in the fractions of Lycium shawii. Major constituents from GC/MS, were 9, 12-octadecadienoic acid (Z, Z), methyl ester, 2,7-Octadiene-1,6-diol and 2,6-dimethyl hydroxy linalool but further five compounds (i.e. lupenone, betulin, lupeol acetate, stigmasterol and β-sitosterol-D-glucoside) were isolated and identified. Treatment of rats with the seeds extract post acrylamide administration ameliorated female sex hormones, oxidative stress, inflammation, DNA damage, and ovarian structure. In conclusion, Lycium shawii petroleum ether seeds fraction may be considered as a nutraceutical agent for improving infertility disorders, oxidative stress and inflammation due to its richness with biologically active phenolic and flavonoids compounds.

Ameliorating effects of adropin on letrozole-induced polycystic ovary syndrome via regulating steroidogenesis and the microbiota inflammatory axis in rats.

Growing evidence supports the role of gut microbiota in chronic inflammation, insulin resistance (IR) and sex hormone production in polycystic ovary syndrome (PCOS). Adropin plays a pivotal role in the regulation of glucose and lipid metabolism and is negatively correlated with IR, which affects intestinal microbiota and sex hormones. However, the effect of adropin administration in PCOS has yet to be investigated. The present study aimed to assess the effects of adropin on letrozole (LTZ)-induced PCOS in rats and the potential underlying mechanisms. The experimental groups were normal, adropin, letrozole and LTZ+adropin. At the end of the experiment, adropin significantly ameliorated PCOS, as evidenced by restoring the normal ovarian structure, decreasing the theca cell thickness in antral follicles, as well as serum testosterone and luteinizing hormone levels and luteinizing hormone/follicle-stimulating hormone ratios, at the same time as increasing granulosa cell thickness in antral follicles, oestradiol and follicle-stimulating hormone levels. The ameliorating effect could be attributed to its effect on sex hormone-binding globulin, key steroidogenic genes STAR and CYP11A1, IR, lipid profile, gut microbiota metabolites-brain-ovary axis components (short chain fatty acids, free fatty acid receptor 3 and peptide YY), intestinal permeability marker (zonulin and tight junction protein claudin-1), lipopolysaccharides/Toll-like receptor 4/nuclear factor kappa B inflammatory pathway and oxidative stress makers (malondialdehyde and total antioxidant capacity). In conclusion, adropin has a promising therapeutic effect on PCOS by regulating steroidogenesis, IR, lipid profile, the gut microbiota inflammatory axis and redox homeostasis. KEY POINTS: Adropin treatment reversed endocrine and ovarian morphology disorders in polycystic ovary syndrome (PCOS). Adropin regulated the ovarian steroidogenesis and sex hormone-binding globulin in PCOS. Adropin improved lipid profile and decreased insulin resistance in PCOS. Adropin modulated the components of the gut-brain-ovary axis (short chain fatty acids, free fatty acid receptor 3 and peptide YY) in PCOS. Adropin improved intestinal barrier integrity, suppressed of lipopolysaccharides/Toll-like receptor 4/nuclear factor kappa B signalling pathway and oxidative stress in PCOS.

P-589 Effects of phthalates on the 3D spheroid cell culture as a model for the human ovarian extracellular matrix

Abstract Study question Do phthalates affect the ovary by impacting its extracellular matrix (ECM)? Summary answer Phthalate mixture (PM) negatively affects the components of ovarian ECM, as demonstrated by spheroids as a model. What is known already The widespread production and pervasive use of phthalates as plasticizers in everyday products significantly increases human exposure to these substances. Their documented toxic effects on reproductive and endocrine functions raise substantial public health concerns. Yet, a full understanding of how phthalates affect ovarian functions and especially the ECM structure in females—more exposed to these products than men—remains incomplete. Moreover, there has been limited investigation in identifying a suitable model capable of mimicking the 3D ovarian ECM structure in vitro for studying phthalate impacts. Study design, size, duration Stromal cells were isolated from the ovarian cortex of post-menopausal organ donors. Cells were seeded onto agarose micro-well molds to form the spheroids. After 6 days, spheroids were treated with a PM (0 and 100 µg/ml) for 4 days. Cell proliferation, spheroids diameter and protein levels of ECM components were measured for both groups. Participants/materials, setting, methods Ovaries from four deceased post-menopausal multi-organ donors (IRB reference 2018/19DEC/475) were used for cell isolation. After seeding of cells, their diameter was measured on days 6 (start of treatment) and 10 (end of treatment) using an inverted microscope to assess their development. The study measured the protein levels of collagen VI, Emilin, and Fibrilin using immunohistochemistry and Ki67 using immunofluorescence in ovarian spheroids. The picrosirius red staining (PSR) was used to measure the collagen content. Main results and the role of chance The evaluation of spheroid diameter indicated comparable sizes on day 6 for both groups (0 µg/ml PM: 109±1.3 µm; 100 µg/ml PM: 112.5±1.1 µm). By day 10, there were no notable differences in spheroid size for the group exposed to PM in comparison to the control group (0 µg/ml PM: 104±1.2 µm; 100 µg/ml PM: 107.2±1.2 µm). The assessment of proliferation revealed no significant difference in the expression of ki67-positive cells between the groups (0 µg/ml PM: 3.211±0.64 %; 100 µg/ml PM: 3.379±0.61%; Fig 1). On the other hand, immunostaining results displayed a decrease in EMILIN-1 (171.6±5.31 vs 201.9±4.71 in control, P &amp;lt; 0.0001) and fibrillin levels (53.1±3.34 vs. 88.3±3.68 in control, P &amp;lt; 0.0001) post-PM treatment. Conversely, collagen VI levels were significantly higher in spheroids treated with PM compared to the control group (122.8±4.99 vs 104.8±4.04, P = 0.005). Although PSR revealed an increase following treatment (9.62±0.75 vs 8.64±0.70 in control), this change did not reach statistical significance (P = 0.344). Limitations, reasons for caution This study exclusively focused on ovarian stromal cells from post-menopausal women. Further research is needed to explore the impacts of PM on ECM of prepubertal and reproductive-age women. Wider implications of the findings Our study highlights the detrimental impact of phthalates on the ECM of postmenopausal women. Findings suggest ovarian stiffness by decreased elastogenesis-associated proteins (EMILIN-1, fibrillin-1) and increased collagen VI (not I, III) post-phthalate exposure.This pioneering study prompts further exploration of phthalate-induced ECM changes and their potential influence on steroidogenesis and folliculogenesis. Trial registration number not applicable

The Protective Effect of Gallic Acid Against Bisphenol A-Induced Ovarian Toxicity and Endocrine Disruption in Female Rats.

Purpose: This study aimed to investigate the protective effects of gallic acid (GA) against ovarian damage induced by bisphenol A (BPA) exposure in female rats. We evaluated whether GA can mitigate the adverse effects of BPA on ovarian structure, inflammatory markers, oxidative stress, apoptosis, and reproductive hormone levels. Methods: Thirty-two female rats were categorized into four groups: control, GA, BPA, and GA+BPA. Histopathological evaluations of ovarian tissue were performed using hematoxylin-eosin staining. The immunohistochemical analysis was conducted for inflammatory, oxidative DNA damage, and apoptotic markers (Tumor necrosis factor alpha [TNFα], cyclooxygenase-2 [COX2], interleukin-1 beta [IL-1β], 8-hydroxydeoxyguanosine [8-OHdG], and caspase 3). Oxidative stress was assessed by measuring malondialdehyde and superoxide dismutase levels. Furthermore, follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrogen, and progesterone levels were quantified using enzyme-linked immunosorbent assay. Results: Histopathological outcomes revealed that BPA significantly induced follicular degeneration, which was effectively mitigated by GA treatment (P < 0.05). Immunohistochemical analysis highlighted the exacerbation of inflammatory responses and oxidative DNA damage and apoptosis (TNFα, COX-2, IL-1β, 8-OHdG, and caspase 3) in BPA-exposed tissues, which were reduced in the presence of GA (P < 0.05). The assessment of oxidative stress demonstrated that GA could significantly decrease lipid peroxidation and partially restore antioxidant defense mechanisms disrupted by BPA (P < 0.05). Hormonal profiling indicated that BPA exposure altered the levels of FSH, LH, estrogen, and progesterone, with GA treatment showing a capacity to modulate these changes, especially in progesterone levels (P < 0.05). Conclusions: The findings suggest that GA exhibits protective properties against BPA-induced ovarian damage through its antioxidative and anti-inflammatory activities, alongside its ability to modulate hormonal imbalances. This research underscores the therapeutic potential of GA in safeguarding reproductive health against environmental toxicants.

Discovery and evaluation of novel spiroheterocyclic protective agents via a SIRT1 upregulation mechanism in cisplatin-induced premature ovarian failure

Currently, no effective treatment exists for premature ovarian failure (POF). To obtain compounds with protective effects against POF, we aimed to design and synthesize a series of spiroheterocyclic protective agents with a focus on minimizing toxicity while enhancing their protective effect against cisplatin-induced POF. This was achieved through systematic modifications of Michael receptors and linkers within the molecular structure of 1,5-diphenylpenta-1,4-dien-3-one analogs. To assess the cytotoxicity and activity of these compounds, we constructed quantitative conformational relationship models using an artificial intelligence random forest algorithm, resulting in R2 values exceeding 0.87. Among these compounds, j2 exhibited optimal protective activity. It significantly increased the survival of cisplatin-injured ovarian granulosa KGN cells, improved post-injury cell morphology, reduced apoptosis, and enhanced cellular estradiol (E2) levels. Subsequent investigations revealed that j2 may exert its protective effect via a novel mechanism involving the activation of the SIRT1/AKT signal pathway. Furthermore, in cisplatin-injured POF in rats, j2 was effective in increasing body, ovarian, and uterine weights, elevating the number of follicles at all levels in the ovary, improving ovarian and uterine structures, and increasing serum E2 levels in rats with cisplatin-injured POF. In conclusion, this study introduces a promising compound j2 and a novel target SIRT1 with substantial protective activity against cisplatin-induced POF.

Strategies for developing 3D printed ovarian model for restoring fertility.

Ovaries play a crucial role in the regulation of numerous essential processes that occur within the intricate framework of female physiology. They are entrusted with the responsibility of both generating a new life and orchestrating a delicate hormonal symphony. Understanding their functioning is crucial for gaining insight into the complexities of reproduction, health, and fertility. In addition, ovaries secrete hormones that are crucial for both secondary sexual characteristics and the maintenance of overall health. A three-dimensional (3D) prosthetic ovary has the potential to restore ovarian function and preserve fertility in younger females who have undergone ovariectomies or are afflicted with ovarian malfunction. Clinical studies have not yet commenced, and the production of 3D ovarian tissue for human implantation is still in the research phase. The main challenges faced while creating a 3D ovary for invivo implantation include sustenance of ovarian follicles, achieving vascular infiltration into the host tissue, and restoring hormone circulation. The complex ovarian microenvironment that is compartmentalized and rigid makes the biomimicking of the 3D ovary challenging in terms of biomaterial selection and bioink composition. The successful restoration of these properties in animal models has led to expectations for the development of human ovaries for implantation. This review article summarizes and evaluates the optimal 3D models of ovarian structures and their safety and efficacy concerns to provide concrete suggestions for future research.